CN113209312A

CN113209312A - Application of reagent for inhibiting expression of transcription factor MEF2C in preparation of medicine for treating keloid

Info

Publication number: CN113209312A
Application number: CN202110490632.0A
Authority: CN
Inventors: 池光范; 李香兰; 姜日花; 孙英淑
Original assignee: Jilin University
Current assignee: Jilin University
Priority date: 2021-05-06
Filing date: 2021-05-06
Publication date: 2021-08-06
Anticipated expiration: 2041-05-06
Also published as: CN113209312B

Abstract

The invention provides application of a reagent for inhibiting expression of a transcription factor MEF2C in preparation of a medicament for treating keloid, and relates to the technical field of skin injury repair. The invention effectively reduces the expression of I type collagen and III type collagen synthesized by keloid fibroblast by inhibiting the expression of transcription factor MEF2C gene, reduces the deposition of extracellular matrix and improves keloid tissue. The invention also provides a keloid fibroblast cell line expressed by the low MEF2C and a medicament for treating keloids.

Description

Application of reagent for inhibiting expression of transcription factor MEF2C in preparation of medicine for treating keloid

Technical Field

The invention belongs to the technical field of skin injury repair, and particularly relates to application of a reagent for inhibiting expression of a transcription factor MEF2C in preparation of a medicament for treating keloid.

Background

Keloid (keloid) is a skin fibroblast cell abnormal proliferation disease related to genetic and immune functions, and is characterized by the pathological condition of fibroblast cell hyperproliferation and excessive deposition of collagen-based Extracellular matrix (ECM) components, and is also considered to be a benign connective tissue tumor of skin. It is a pathological wound healing response secondary to skin injury such as trauma, burn, infection, acne, folliculitis, perforation of the ear or surgical wound. Unlike hyperplastic scars which are limited in the original wound area, keloid lesion obviously exceeds the original injury range, the edge of the keloid lesion is always in a crab foot shape, the keloid lesion grows invasively to the surrounding normal tissues, the keloid lesion does not degenerate all the time, and the keloid lesion is not recurrent after simple operation treatment. Keloid is a unique disease of human, the incidence of the keloid is different in species, the incidence of the keloid is higher in black and yellow race, the incidence of the keloid is as high as 4-16% in yellow race, and the keloid is common to young people and has family genetic susceptibility. The disease is an intractable skin disease, which not only affects the external image of a patient, but also is accompanied by pruritus and pain, seriously affects the life quality of the patient, and can also affect the motor function if the skin is greatly damaged. So far, no thorough cure method exists, the treatment is mainly symptomatic treatment, although the treatment methods are more, the radical cure can not be achieved, and the relapse is easy. Therefore, the molecular pathogenesis of the keloid is clarified, and research and development of a new medicine for treating the keloid are focused on the specific links of the keloid.

Disclosure of Invention

In view of the above, the present invention aims to provide an application of a reagent for inhibiting expression of transcription factor MEF2C in preparation of a drug for treating keloid, wherein transcription factor MEF2C is used as a biomarker for keloid, and through targeting specific inhibition of transcription factor MEF2C highly expressed in keloid, synthesis and secretion of type I collagen extracellular matrix and type III collagen extracellular matrix by fibroblasts are inhibited, and downstream target genes regulated by MEF2C are controlled, so as to achieve the purpose of controlling and treating keloid.

In order to achieve the above object, the present invention provides the following technical solutions:

the invention provides application of a reagent for inhibiting expression of a transcription factor MEF2C in preparation of a medicament for treating keloid.

Preferably, the agent for inhibiting expression of transcription factor MEF2C comprises interfering RNA of double-stranded MEF2C or lentivirus carrying an interfering gene interfering expression of MEF 2C.

Preferably, the interfering RNA of the double-stranded MEF2C is used for inhibiting the expression of MEF2C target gene of keloid cells and inhibiting the expression of MEF2C protein in a transient transfection method.

Preferably, the nucleotide sequence of the interfering RNA of double-stranded MEF2C comprises MEF2C siRNA-1 or MEF2C siRNA-2; the nucleotide sequence of the MEF2C siRNA-1 is shown in SEQ ID NO.1, and the nucleotide sequence of the MEF2C siRNA-2 is shown in SEQ ID NO. 2.

Preferably, the lentivirus carrying the interference gene interfering with the expression of MEF2C is used for inhibiting the expression of MEF2C and the expression of MEF2C as target genes of keloid cells in a stable transfection method.

Preferably, the keloid cells comprise keloid fibroblasts.

The invention also provides a keloid fibroblast cell line with low expression of MEF 2C.

The invention also provides a construction method of the cell line, which is characterized in that a recombinant expression vector is introduced into the keloid fibroblast, and the sequence of the recombinant expression vector contains the interfering RNA of double-stranded MEF 2C; the nucleotide sequence of the interfering RNA of double-stranded MEF2C comprises MEF2C siRNA-1 or MEF2C siRNA-2; the nucleotide sequence of the MEF2C siRNA-1 is shown in SEQ ID NO.1, and the nucleotide sequence of the MEF2C siRNA-2 is shown in SEQ ID NO. 2.

Preferably, the recombinant expression vector comprises a plasmid vector, a lentiviral vector, an adenoviral vector or a retrovirus.

The invention also provides a medicament for treating keloid, and the effective component of the medicament comprises a reagent for inhibiting the expression of the transcription factor MEF 2C.

The invention provides application of a reagent for inhibiting expression of a transcription factor MEF2C in preparation of a medicament for treating keloid, which effectively reduces expression of I type collagen and III type collagen synthesized by keloid fibroblasts, reduces deposition of extracellular matrix and improves keloid tissues by inhibiting expression of a transcription factor MEF2C gene.

In the invention, after inhibiting the expression of MEF2C gene, the low expression MEF2C keloid fibroblast can inhibit fibroblast from synthesizing and secreting extracellular matrixes such as type I collagen and type II collagen and control downstream target genes regulated and controlled by MEF2C, effectively inhibit and control the characteristics of keloid fibroblast, and achieve the treatment effect of controlling and treating keloid. In the embodiment of the invention, the siRNA (MEF2C siRNA-1 or MEF2C siRNA-2) is utilized to promote the low expression of MEF2C in keloid fibroblasts, which means that the mRNA expression quantity of MEF2C in the keloid fibroblasts is reduced by at least 50%, so that the content of collagen in the keloid is reduced, and the purpose of treating the keloid with reduced volume is achieved.

Drawings

FIG. 1 shows a strong positive expression of MEF2C protein in keloid tissues compared to normal skin tissues, where A is the immunohistochemical staining pattern for normal skin and keloid MEF2C and B is the quantitative analysis of the immunochemical staining results;

FIG. 2 shows that the keloid tissues significantly highly express MEF2C protein compared to normal skin tissues, where A is the Western blot analysis of normal skin and keloid tissues MEF2C protein expression, and B is the quantitative analysis of the Western blot results;

figure 3 shows keloid fibroblasts are spread, bulky and morphologically irregular compared to normal skin fibroblasts, with the left image being a hypo-specular (100 x) and the right image being a hyper-specular (200 x);

FIG. 4 shows that keloid fibroblasts highly express MEF2C (red) and are located in the nucleus (DAPI, blue) compared to normal skin fibroblasts, where A is the MEF2C expression map of keloid fibroblasts observed by cell immunoduplex fluorescent staining and B is the MEF2C positive rate quantification map;

FIG. 5 shows that keloid fibroblasts highly express mRNA of MEF2C, Collagen I and Collagen III, wherein A is mRNA RT-PCR electrophoretogram of MEF2C, Collagen I and Collagen III, and B is electrophoresis result semiquantitative analysis graph, compared with normal fibroblasts;

FIG. 6 shows that 5ng/ml, 10ng/ml and 20ng/ml of TGF-beta 1 all significantly improve the expression of the keloid fibroblast MEF2C, Collagen I and Collagen III proteins, wherein A is the result of Western blot analysis on the influence of different concentrations of TGF-beta 1 on the expression of proteins such as keloid fibroblast MEF2C, B, C and D are the results of quantitative analysis on the change of expression levels of MEF2C, Collagen I and Collagen III proteins under the stimulation of different concentrations of TGF-beta 1;

FIG. 7 shows that all 3 MEF2C siRNAs had interference effects on MEF2C, but of these, MEF2C siRNA-1 and MEF2C siRNA-2 had the most significant effect on inhibiting the expression of Collagen I and Collagen III proteins, where A is the effect on the expression of Collagen I and Collagen III proteins after Western blot analysis that siRNA interferes with the keloid fibroblast MEF2C protein; B. effect of siRNA with C and D being 3 MEF2C, respectively, on MEF2C, Collagen I and Collagen III protein expression.

Detailed Description

The reagent for inhibiting the expression of the transcription factor MEF2C preferably comprises double-stranded MEF2C interfering RNA or interfering gene lentivirus carrying interfering MEF2C expression. In the present invention, using the interfering RNA of said double-stranded MEF2C, it is preferable to suppress expression of MEF2C, a target gene of keloid cells, and to suppress expression of MEF2C protein, in a transient transfection method; and the nucleotide sequence of the interfering RNA of double-stranded MEF2C preferably comprises MEF2C siRNA-1 or MEF2C siRNA-2; the nucleotide sequence of MEF2C siRNA-1 is shown in SEQ ID NO.1 (5'-CATGCGACTTTCTGAAGGA-3'), and the nucleotide sequence of MEF2C siRNA-2 is shown in SEQ ID NO.2 (5'-GCTGTAGCAGTTCGTACGA-3').

In the invention, the keloid mainly comprises cellular fibroblast high-expression MEF2C, and after the interference of the interfering RNA, the expression quantity of the keloid fibroblast MEF2C can be obviously inhibited, the content of collagen in the keloid is reduced, and the treatment purpose of reducing the volume of the keloid is achieved. The method of transient transfection is not particularly limited in the present invention, and any conventional transient transfection method in the art may be used.

The invention utilizes the interference gene lentivirus carrying interference MEF2C expression, preferably inhibits the expression of target gene MEF2C of keloid cells and inhibits the expression of MEF2C protein by a stable transfection method. The keloid cells of the present invention include keloid fibroblasts.

The invention also provides a keloid fibroblast cell line with low expression of MEF 2C. The invention can obtain the keloid fibroblast with low expression MEF2C by utilizing the interfering RNA of the double-chain MEF2C or interfering gene lentivirus carrying the expression of interfering MEF2C or other viruses to inhibit the expression of the target gene MEF2C of the keloid fibroblast. The cell line of the invention reduces the expression level of MEF2C in fibroblasts, modifies the cytology characteristics of keloid fibroblasts, reduces the expression of extracellular matrix related genes (Collagen I and Collagen III), and simultaneously can inhibit the over-synthesis of Collagen, thereby achieving the purpose of treating keloid.

The type of the recombinant expression vector is not particularly limited in the present invention, and any type capable of introducing siRNA or nucleotide sequence into a vector may be used, and preferably includes a plasmid vector, a lentiviral vector, an adenoviral vector, or a retrovirus. The method for constructing the recombinant expression vector of the present invention is not particularly limited, and a conventional construction method in the art may be used.

The invention also provides a medicament for treating keloid, and the effective component of the medicament comprises a reagent for inhibiting the expression of the transcription factor MEF 2C. The agent for inhibiting the expression of the transcription factor MEF2C according to the present invention is preferably the same as described above, and will not be described herein. The dosage form of the medicine of the invention preferably comprises injection, injection or skin sticking film agent.

The invention also provides a method for treating keloid, which comprises the step of enabling a reagent (the effective component is the interfering RNA of double-chain MEF 2C) for inhibiting the expression of the transcription factor MEF2C to absorb the reagent at the keloid part in a subcutaneous injection, intravenous injection or skin sticking film mode, thereby achieving the purpose of treating the keloid.

The following examples are provided to illustrate the application of the agent for inhibiting expression of the transcription factor MEF2C in the preparation of a medicament for treating keloid, but they should not be construed as limiting the scope of the present invention.

Example 1

Differential expression of MEF2C protein in human keloid tissues

1.1 collect 20 keloid tissue samples, including 10 keloid scars on the chest and back and 10 keloid scars on the ears. Keloid tissue samples were all from the dermatology department of the midday friendship hospital, the university of Jilin. The obtained tissue is fixed by formalin to prepare a wax block tissue specimen and a tissue section, and is subjected to tissue immunochemical staining.

The immunohistochemical method comprises the following steps: steps 4-5, 8-9 Using UltraSensitive^TMSP (Mouse/Rabbit) IHC immunohistochemical KIT (McXin Biotechnology development Co., Ltd., KIT-9710) was performed according to the instructions.

(1) The thickness of the paraffin tissue section is 4 mu m;

(2) slicing, dewaxing and hydrating according to conventional steps;

(3) adopting a high-temperature high-pressure method to repair the tissue antigen;

(4) blocking endogenous peroxidase with 3% hydrogen peroxide;

(5) blocking with 10% BSA;

(6) plus primary antibody (MEF2C, 1:200, Abcam);

(7) reacting at 4 ℃ in a refrigerator overnight;

(8) reacting with horse radish enzyme labeled goat anti-mouse/rabbit IgG polymer;

(9) DAB color development;

(10) counter staining with hematoxylin and transparentizing;

(11) sealing neutral gum into a sheet;

(12) and (5) photographing and analyzing the result.

In immunohistochemical staining, both keloid tissues highly expressed MEF2C, which was statistically significant compared to normal skin tissues (fig. 1).

1.2 extracting protein from scar tissue of human body, and comparing and analyzing expression of MEF2C protein in normal skin tissue and keloid tissue by Western Blot method

The method is carried out according to a conventional Western Blot method, and the steps are briefly described as follows: (1) protein extraction: the tissue was placed in a mill and milled by pouring in appropriate liquid nitrogen. According to the PMSF: preparing lysate in a ratio of RIPA to 100, adding 300-500 mu l of lysate into mucosal tissue, and fully lysing the mucosal tissue on ice; cracking on ice for 30 min; precooling a low-temperature centrifuge in advance, placing the collected liquid in the centrifuge, and centrifuging for 15min at 12000rmp and 4 ℃; the supernatant was collected in an EP tube and stored at-80 ℃.

(2) The BCA method is used for drawing a standard protein curve and measuring the protein concentration.

(3) Preparing glue: according to the conventional formula, separating gel (10% concentration, 10ml) and concentrating gel (5% concentration, 3ml) are prepared.

(4) Protein loading: protein samples were added with 5 Xprotein loading buffer and boiled for 5min, and 40. mu.g protein was loaded per well.

(5) Protein electrophoresis concentration, separation and mold conversion are carried out, and the mold is sealed by 5 percent of skimmed milk powder after mold conversion.

(6) Primary antibody incubation: MEF2C (1: 2000, Abcam), Collagen I (1:5000, Abcam), Collagen III (1:5000, Abcam), at 4 ℃ overnight.

(7) And (3) secondary antibody incubation: diluting a secondary antibody (1:5000, goat anti-rabbit Ig G-HRP antibody, China Beijing China fir Jinqiao biological Co., Ltd.), incubating with a membrane for color development, reacting with ECL color development solution for color development, and taking a picture in a gel image analyzer.

The results showed that scar tissue significantly highly expressed MEF2C protein (P < 0.01) compared to normal tissue (fig. 2).

Example 2

2.1 isolated culture of patient keloid fibroblast

The keloid fibroblast and normal skin fibroblast are separated and cultured by a type I collagenase digestion method. The general steps are as follows:

(1) placing keloid or normal skin tissue cut off in the operation into a centrifuge tube containing sterile buffer solution, and repeatedly rinsing for 3-5 times until the liquid is clear. For keloid tissues with a diameter greater than 2cm, only the tissue of the edge region within 0.5cm from the edge of the scar was left for cell culture.

(2) The tissue was transferred to a sterile petri dish and the subcutaneous adipose tissue was removed with an ophthalmic scissors. A portion of the deep dermis of the keloid scar was removed (about 1/4 for the entire dermis), and the entire dermis of normal skin was retained.

(3) The tissue is cut into strips with a width of about 1-2 mm, 0.2% neutral protease solution is added, and digestion is carried out overnight at 4 ℃.

After overnight, the epidermis and dermis were torn apart with ophthalmologic forceps, and the dermis was repeatedly cut to a paste, 10 volumes of 0.2% collagenase I solution was added, and the mixture was subjected to shake digestion at 37 ℃ for 3-5 hours until large tissue masses disappeared.

(4) The digestion was stopped by the addition of an equal amount of complete medium and the aspiration was repeated up and down with a pipette to free the cells entrapped in the incompletely digested fibers. The liquid was filtered through a 80 mesh sieve to remove the incompletely digested tissue.

(5) The filtrate 1200g was centrifuged for 5 min. The supernatant was discarded, the pellet was retained, and the cells were resuspended by adding complete medium. After cell counting, the number of cells was 1X 10⁵The cells were inoculated at a density of one/ml into a culture flask and placed at 37 ℃ with 5% CO₂Cultured in an incubator. The liquid is changed for the first time after 5 days, and the liquid is changed once every day later. If not otherwise stated, cells were cultured in complete medium and after complete cell growth to complete confluence were digested with trypsin solution at a rate of 1: passage 2.

(6) When the cells passed to passage 3, various tests and treatments were performed. The morphology of the keloid fibroblasts was identified by inverted phase contrast microscopy.

The morphology of keloid fibroblasts, shown by inverted phase contrast microscopy, was hypertrophic and significantly larger in volume than normal fibroblasts (fig. 3). The cellular immunofluorescence identification result shows that MEF2C is remarkably and highly expressed in the nucleus of the keloid fibroblast cells cultured in an isolation way (figure 4).

2.2 comparison of mRNA expression levels of MEF2C in keloid fibroblasts and normal skin fibroblasts

Extracting total RNA of keloid fibroblasts and normal skin fibroblasts by an RNA extraction kit, carrying out reverse transcription to obtain cDNA, and detecting mRNA expression of MEF2C, Collagen I and Collagen III of the fibroblasts by PCR.

Extracting total RNA of fibroblast (keloid fibroblast, normal skin fibroblast) by using an RNA extraction kit, and performing reverse transcription to obtain cDNA, wherein the reverse transcription system is as follows: 500ng of total RNA, 1. mu.l of Oligo-thymine 18primer (adsorbed Oligo (dT)18primer) (0.5. mu.g/. mu.l), 10. mu.l of 2 XTS Reaction Mix (TS Reaction Mix), 1. mu.l of transporter Mix (ransScript RT/RI Enzyme Mix) T and 1. mu.l of gDNA remover (gDNAROMOVER) were supplemented to 20. mu.l with double distilled water. The reverse transcription reaction temperature was set to: reacting at 42 ℃ for 15 minutes and 85 ℃ for 5 minutes, and gradually cooling to 4 ℃ after the reaction is stopped.

Each of the groups of MEF2C, Collagen I and Collagen III was tested for expression by PCR using primers synthesized by Shanghai bioengineering, Inc. The primer sequences are as follows:

MEF2C-R(SEQ ID NO.3)：5’-CGAGATGCCAGTCTCCATCC-3’，

MEF2C-F(SEQ ID NO.4)：5’-AGCAGACCTGGTGAGTTTCG-3’；

Collagen I-R(SEQ ID NO.5)：5’-GGTGGTGGTTATGACTTTGG-3’，

Collagen I-F(SEQ ID NO.6)：5’-GTTCTTGGCTGGGATGTTTT-3’；

Collage III-R(SEQ ID NO.7)：5’-GCTCTGCTTCATCCCACTATTA-3’，

Collage III-F(SEQ ID NO.8)：5:’-TGCGAGTCCTCCTACTGCTAC-3’；

GAPDH-R(SEQ ID NO.9)：5’-CGCTCTCTGCTCCTCCTGTT-3’，

GAPDH-F(SEQ ID NO.10)：5’-CCATGGTGTCTGAGCGATGT-3’。

the reaction system is as follows: 10 μ l of Green Master mix polymerase (u:)

Green Master Mix 2X), 0.5. mu.l of the forward primer (upflow primer, 10. mu.M), 0.5. mu.l of the reverse primer (downstream primer, 10. mu.M), 1000ng of cDNA, the system was made up to 25. mu.l with double distilled water. The reaction temperature was set as: pre-denaturation reaction at 95 ℃ for 2 minutes, and then 30-40 cycles of 30 seconds of denaturation reaction at 95 ℃, 30 seconds of renaturation reaction at 56 ℃ and 30 seconds of extension reaction at 72 ℃ in one cycle.

Protein and mRNA were extracted from the first or second generation keloid fibroblasts and normal skin fibroblasts, and it was found that the keloid fibroblasts significantly highly expressed mRNA and corresponding protein (P < 0.01) of MEF2C, Collagen I and Collagen III, compared to normal tissue (FIG. 5).

Example 3

Keloid fibroblasts up-regulated the expression of MEF2C, Collagen I and Collagen III proteins following TGF- β 1 stimulation.

The collected keloid fibroblasts from passage 2 to passage 4 were used to observe the effect of TGF- β 1 on the expression of the keloid fibroblast MEF2C, Collagen I and Collagen III proteins. TGF-beta 1 treatment concentrations were 1ng/ml, 2.5ng/ml, 5ng/ml, 10ng/ml and 20ng/ml, respectively. Western blot procedure reference example 2.

As shown in FIG. 6, the protein levels of MEF2C, Collagen I and Collagen III of keloid fibroblasts increased to various degrees under the induction of TGF-. beta.1, and were most significantly changed at TGF-. beta.15 ng/ml (P < 0.01). Experimental results show that TGF-beta 1 promotes the synthesis of keloid fibroblast MEF2C, Collagen I and Collagen III proteins.

Example 4

Transfection of siRNA into keloid fibroblasts MEF2C siRNA was analyzed for the effect on fibroblast Collagen I and Collagen III protein expression.

The effective siRNA of MEF2C is: MEF2C siRNA-1(SEQ ID NO. 1): 5'-CATGCGACTTTCTGAAGGA-3' the flow of the air in the air conditioner,

MEF2C siRNA-2(SEQ ID NO.2)：5’-GCTGTAGCAGTTCGTACGA-3’，

MEF2C siRNA-3(SEQ ID NO. 11): 5'-CGTAGCAACTCCTACTTTA-3' are provided. The siRNA was manufactured by Eton Biotechnology, Inc., Guangzhou, Inc., under product numbers stB0007066A, stB0007066B and stB 0007066C.

And (3) transfection step: transfection concentration of siRNA: the MEF2C siRNA transfection concentration was 50 nM.

2. Transfection with lipofectamine2000(lipo2000, Invitrogen): (1) one day before transfection, 5X 10 inoculation⁴And (3) putting each cell/ml into a cell culture plate, so that the cell density during transfection can reach 30-50%.

(2) For each transfection sample, siRNA-lipo2000 mixtures were prepared as follows: a. dilution of transfection reagent lipo 2000: before use, the lipo2000 transfection reagent is shaken up lightly, then an appropriate amount is taken, diluted by serum-free optimized medium (Opti-MEMI), mixed lightly and incubated for 5min at room temperature; b. and (3) diluting siRNA: the siRNA was diluted with Opti-MEMI without serum and mixed gently; c. after 5min incubation, the diluted lipo2000 was gently mixed with the diluted siRNA of (b) above, and incubated at room temperature for 20min to form siRNA-lipo2000 mixture, which may be cloudy, but does not affect transfection.

(3) The siRNA-lipo2000 mixture was added to a cell culture plate containing cells and culture medium and mixed by gentle shaking.

(4) Place the plates in 37 ℃ CO₂And culturing in an incubator for 24-48 h until detection time.

(5) And (4) detecting the protein level.

The results are shown in FIG. 7, which shows that both MEF2C siRNA-1 and MEF2C siRNA-2 have different degrees of inhibition effects on the expression of the Collagen I and Collagen III proteins, which are extracellular matrices of keloid fibroblasts, and especially the MEF2C siRNA-2 inhibition effect is more significant (P < 0.01).

The above examples show that keloid mainly comprises cellular fibroblast high-expression MEF2C, and the expression of keloid fibroblast MEF2C is significantly inhibited by applying siRNA interference of MEF2C, so that the content of collagen in keloid can be reduced, and the purpose of treating keloid volume reduction is achieved. The results show that the siRNA and the like can inhibit the synthesis of collagen of keloid fibroblasts by interfering the expression of MEF2C protein, and the keloid treatment effect is effectively promoted.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

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Claims

1. Application of a reagent for inhibiting expression of transcription factor MEF2C in preparation of a medicament for treating keloid is provided.

2. The use of claim 1, wherein the agent that inhibits the expression of the transcription factor MEF2C comprises interfering RNA of double-stranded MEF2C or a lentivirus carrying an interfering gene that interferes with the expression of MEF 2C.

3. The use according to claim 2, wherein said double-stranded interfering RNA of MEF2C is used to suppress expression of MEF2C, a target gene of keloid cells, and to suppress expression of MEF2C protein, in a transient transfection process.

4. The use of claim 2 or 3, wherein the interfering RNA of double-stranded MEF2C comprises MEF2C siRNA-1 or MEF2C siRNA-2; the nucleotide sequence of the MEF2C siRNA-1 is shown in SEQ ID NO.1, and the nucleotide sequence of the MEF2C siRNA-2 is shown in SEQ ID NO. 2.

5. The use according to claim 2, wherein said lentivirus carrying an interfering gene interfering with expression of MEF2C is used to suppress expression of MEF2C, a target gene of keloid cells, and to suppress expression of MEF2C protein, in a stable transfection procedure.

6. The use of claim 3 or 5, wherein said keloid cells comprise keloid fibroblasts.

7. A cell line comprising keloid fibroblasts that underexpress MEF 2C.

8. The method of claim 7, wherein a recombinant expression vector comprising the interfering RNA of double-stranded MEF2C in its sequence is introduced into keloid fibroblasts; the interfering RNA of the double-stranded MEF2C comprises MEF2C siRNA-1 or MEF2C siRNA-2; the nucleotide sequence of the MEF2C siRNA-1 is shown in SEQ ID NO.1, and the nucleotide sequence of the MEF2C siRNA-2 is shown in SEQ ID NO. 2.

9. The method of claim 8, wherein the recombinant expression vector comprises a plasmid vector, a lentiviral vector, an adenoviral vector, or a retrovirus.

10. A medicament for treating keloid characterized in that the active principle of said medicament comprises an agent inhibiting the expression of the transcription factor MEF 2C.